New guinea impatiens variety SAKIMP069

ABSTRACT

A New Guinea Impatiens plant designated SAKIMP069 is disclosed. Embodiments include seeds of New Guinea Impatiens SAKIMP069, plants of New Guinea Impatiens SAKIMP069, to plant parts of New Guinea Impatiens SAKIMP069, and methods for producing an impatiens plant produced by crossing New Guinea Impatiens SAKIMP069 with itself or with another impatiens variety. Embodiments include methods for producing an impatiens plant containing in its genetic material one or more genes or transgenes and transgenic impatiens plants and plant parts produced by those methods. Embodiments also relate to impatiens varieties, breeding varieties, plant parts, and cells derived from New Guinea Impatiens SAKIMP069, methods for producing other impatiens lines or plant parts derived from New Guinea Impatiens SAKIMP069, and the impatiens plants, varieties, and their parts derived from use of those methods. Embodiments further include hybrid impatiens seeds, plants, and plant parts produced by crossing New Guinea Impatiens SAKIMP069 with another impatiens variety.

BACKGROUND

All publications cited in this application are herein incorporated byreference. New Guinea Impatiens is a species of flowering plants in thefamily Balsaminaeceae.

New Guinea Impatiens can be propagated from seed, cuttings, and tissueculture. Seed, cuttings and tissue culture germination protocols for NewGuinea Impatiens are well-known in the art.

New Guinea Impatiens is an important and valuable ornamental plant.Thus, a continuing goal of ornamental plant breeders is to developplants with novel characteristics, such as color, growth habit, andhardiness. To accomplish this goal, the breeder must select and developplants that have traits that result in superior New Guinea Impatiensvarieties.

The foregoing examples of the related art and limitations relatedtherewith are intended to be illustrative and not exclusive. Otherlimitations of the related art will become apparent to those of skill inthe art upon a reading of the specification.

SUMMARY

The following embodiments and aspects thereof are described inconjunction with systems, tools and methods which are meant to beexemplary, not limiting in scope.

According to one embodiment, there is provided a New Guinea Impatiensplant which is valued as breeding line enabling the development ofsuperior ornamental New Guinea Impatiens plants.

Another embodiment discloses a New Guinea Impatiens plant, wherein asample of representative sample of plant tissue of said New GuineaImpatiens is deposited with a Budapest Depository.

Another embodiment relates to tissue culture produced from protoplastsor cells from the New Guinea Impatiens plants disclosed in the subjectapplication, wherein said cells or protoplasts are produced from a plantpart selected from the group consisting of pollen, ovules, embryos,protoplasts, meristematic cells, callus, leaves, anthers, cotyledons,hypocotyl, pistils, roots, root tips, flowers, seeds, petiole, andstems.

Another embodiment relates to a plant, or a part thereof, produced bygrowing New Guinea Impatiens SAKIMP069, wherein the plant part comprisesat least one cell of New Guinea Impatiens SAKIMP069.

Another embodiment relates to a tissue or cell culture of regenerablecells produced from the plant of SAKIMP069 and a New Guinea Impatiensplant regenerated from the tissue or cell culture of SAKIMP069.

Another embodiment relates to a method of vegetatively propagating theplant of SAKIMP069, comprising the steps of: collecting tissue or cellscapable of being propagated from a plant of SAKIMP069; cultivating saidtissue or cells to obtain proliferated shoots; and rooting saidproliferated shoots to obtain rooted plantlets; or cultivating saidtissue or cells to obtain proliferated shoots, or to obtain plantletsand a plant produced by growing the plantlets or proliferated shoots ofsaid plant.

A further embodiment relates to a method for producing a New GuineaImpatiens seed or embryo, wherein the method comprises crossing aSAKIMP069 plant with a different New Guinea Impatiens plant andharvesting the resultant New Guinea Impatiens seed or embryo.

A further embodiment relates to a method for developing a New GuineaImpatiens plant in a New Guinea Impatiens plant breeding program,comprising applying plant breeding techniques comprising crossing,recurrent selection, mutation breeding, wherein said mutation breedingselects for a mutation that is spontaneous or artificially induced,backcrossing, pedigree breeding, marker enhanced selection,haploid/double haploid production, or transformation to the New GuineaImpatiens plant of SAKIMP069, or its parts, wherein application of saidtechniques results in development of a New Guinea Impatiens plant.

A further embodiment relates to a method of introducing a mutation intothe genome of a New Guinea Impatiens plant SAKIMP069, said methodcomprising mutagenesis of the plant, or plant part thereof, ofSAKIMP069, wherein said mutagenesis is selected from the groupconsisting of temperature, long-term seed storage, tissue cultureconditions, ionizing radiation, chemical mutagens, and targeting inducedlocal lesions in genomes, and wherein the resulting plant comprises atleast one genome mutation and producing plants therefrom.

A further embodiment relates to a method of editing the genome of NewGuinea Impatiens plant SAKIMP069, wherein said method is selected fromthe group comprising zinc finger nucleases, transcription activator-likeeffector nucleases (TALENs), engineered homingendonucleases/meganucleases, and the clustered regularly interspacedshort palindromic repeat (CRISPR)-associated protein9 (Cas9) system, andplants produced therefrom.

A further embodiment relates to a New Guinea Impatiens seed produced bygrowing SAKIMP069.

A further embodiment relates to a method of producing a New GuineaImpatiens plant, or part thereof, by growing a seed produced onSAKIMP069.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by study of thefollowing descriptions.

DETAILED DESCRIPTION

New Guinea Impatiens variety SAKIMP069 disclosed in the presentapplication has shown uniformity and stability, as described in thefollowing section via vegetative cuttings and tissue culture. New GuineaImpatiens variety SAKIMP069 disclosed in the present application hasbeen asexually reproduced a sufficient number of generations withcareful attention to uniformity of plant type and has been increasedwith continued observation for uniformity. Additionally, New GuineaImpatiens variety SAKIMP069 produces viable pollen and is capable ofbeing used as a parental line in breeding programs.

Origin of SAKIMP069

The present invention comprises of a new and distinct cultivar of NewGuinea Impatiens, botanically known as Impatiens×hybrida, and referredto by the variety name ‘SAKIMP069’. SAKIMP069′ originated from aninterspecific hybridization between Impatiens ‘NJ-1174A’, an unpatentedproprietary lilac flowered Impatiens breeding line and Impatiens‘NJ-333-49H-6H’, an unpatented proprietary blush flowered Impatiensbreeding line in Misato, Japan.

In May 2017, the female parent line ‘NJ-1174A’ and male parent line‘NJ-333-49H-6H’ were crossed and a population of F₁ plants was created.The F₁ plants were evaluated in Misato, Japan in an open field trial.The criteria for plant selection included abundance of flowers, purpleflower color, mounding plant habit, and strong root system. At thecompletion of the trial, one single-plant selection was made based onthe above criteria and vegetatively propagated. From June to August2019, the selection was evaluated in an open field in Misato, Japan.Shoot-tip cuttings of the variety were then shipped to Salinas, Calif.,where the plants were regenerated and reevaluated for stability oftraits. The selection subsequently was named ‘SAKIMP069’ and found tohave its unique characteristics reproduce true to type in successivegenerations of asexual propagation.

Environmental Conditions for Plant Growth

The terminal 1.0 to 1.5 inches of an actively growing stem was excised.The vegetative cuttings were propagated in five to six weeks. The baseof the cuttings were dipped for 1 to 2 seconds in a 1:9 solution of DIP'N GROW (1 solution: 9 water) root inducing solution immediately priorto sticking into the cell trays. Cuttings were stuck into plastic celltrays having 98 cells, and containing a moistened peat moss-basedgrowing medium. The cuttings were misted with water from overhead for 10seconds every 30 minutes until sufficient roots were formed.

Rooted cuttings were transplanted and grown in 20 cm diameter plasticpots in a glass greenhouse located in Salinas, Calif. Pots contained apeat moss-based growing medium. Soluble fertilizer containing 20%nitrogen, 10% phosphorus and 20% potassium was applied once a day orevery other day by overhead irrigation. Pots were top-dressed with adry, slow release fertilizer containing 20% nitrogen, 10% phosphorus and18% potassium. The typical average air temperature was 24° C.

Data in Table 1 were taken in Salinas, Calif. Data obtained from plantsgrown 5.25 months from transplant into quart sized pots from rootedcuttings in Salinas, Calif., under greenhouse conditions.

The following traits and characteristics describe the new variety. Colorreferences are to the Royal Horticultural Society Colour Chart, 6thedition. Anatomic labels are from The Cambridge Illustrated Glossary ofBotanical Terms, by M. Hickey and C. King, Cambridge University Press.

TABLE 1 VARIETY DESCRIPTION INFORMATION Characteristic SAKIMP069 Family:Balsaminaceae Species: Impatiens intergeneric hybrid (Impatiens Xhybrida hort) Common Name: Impatiens Parentage Female Parent: NJ-1174AMale Parent: NJ-333-49H-6H General Appearance, Growth, and PropagationPlant Habit: Compact; mounding Plant Height: Approximately 21.0 cm PlantSpread: Approximately 40.0 cm Number of Main Branches: 5-1 center and 4surrounding Length of Main Branches Approximately 3.0 cm to 4.0 cm fromstart to start of secondary branches Diameter of Main BranchesApproximately 1.0 cm Color of Main Branches 187A (Dark Red) and 146A(Moderate Olive Green) Number of Secondary Branches 2 to 3 on each mainbranch Length of Secondary Branches, Approximately 9.0 cm to 14.0 cmexcluding flower(s) Diameter of Secondary Branches Approximately 7.0 mmColor of Secondary Branches 187A (Dark Red) and 146A (Moderate OliveGreen) Life Cycle Annual Time to produce a rooted cutting: About 4 weeksTime to bloom from propagation: 6 to 8 Weeks Flowering requirements(season): Will flower so long as temperature is above 5° C.Resistance/susceptibility: No particular resistance or susceptibilityobserved Temperature tolerances: Plants observed to continue floweringin a temperature range of 5° C. to 36° C. Stems Stem Color: 187A (DarkRed) and 146A (Moderate Olive Green) Stem anthocyanin: 187A (Dark Red)Stem pubescence: None Stem pubescence color: None Stem description:Strong; oval cross - section, smooth and shiny Stem Length:Approximately 8.0 cm from secondary branch to inflorescence StemDiameter: Approximately 4.5 mm Internode Length: Approximately 2.0 cm to5.0 cm Leaves Leaf Arrangement: Whorled with up to 5 leaves per node,opposite if only two leaves at one node Leaf Shape: Lanceolate Leaf Tip:Acuminate Leaf Base: Shortly attenuate Leaf margin: Serrate Leafsurface: Smooth, waxy Leaf Pubescence: None observed Leaf Blade Length:Approximately 7.0 cm for medium leaves Leaf Blade Width: Approximately3.0 cm for medium leaves Leaf Color - Upper: Closest to but much darkerthan NN137A (Greyish Olive Green) Leaf Color - Lower: Background of 191A(Greyish Yellow Green) with specks of 59A (Dark Red) throughout LeafVariegation: None Leaf Fragrance: None Petiole Length: Approximately 8.0mm Petiole Diameter: Approximately 2.0 mm Petiole Color - Upper 144A(Strong Yellow Green) Petiole Color - Lower 183B (Dark Red) PetioleTexture: Slightly pubescent Petiole Pubescence: NN155D (White) LeafVenation Color - Upper: 144A (Strong Yellow Green) midvein Leaf VenationColor - Lower: 183B (Dark Red) midvein Flower Number of Flowers pernode: 1 to 3 in bloom at one time; about 5 to 7 flower buds Number ofFlowers per plant: Approximately 50 Lastingness of Blooms: About 14 daysFlower Inflorescence Type: Single flower with spur Flower Fragrance:Absent Corolla Shape: Roughly circular with 5 radial petals CorollaDiameter: Approximately 5.5 cm to 6.0 cm Corolla Depth: Approximately3.0 mm Sepal Shape: Lanceolate Sepal Number: 2 Sepal Base: SubcordateSepal Apex: Caudate Sepal Margin: Entire Sepal Texture: Glabrous SepalColor - upper Background of N155B (Pinkish White); tip of darker than187A (Dark Red); 146B (Moderate Yellow Green) near tip with specks of59A (Dark Red) throughout Sepal Color - Lower Background of N155B(Pinkish White); tip of darker than 187A (Dark Red); 146B (ModerateYellow Green) near tip with specks of 59A (Dark Red) throughout SepalLength: Approximately 15.0 mm Sepal Width: Approximately 6.0 mm Bud BudSurface Smooth with very slight pubescence on edge Bud LengthApproximately 1.7 cm Bud Diameter Approximately 12.0 mm at midpoint BudShape: Deltoid longitudinal cross-section Bud Color: NN74D (StrongReddish Purple) darkening to 64B (Strong Purplish Red) to 178A (GreyishRed) Peduncle Length Approximately 4.0 cm Peduncle width Approximately2.0 mm Peduncle Texture Lightly pubescent - NN155D (White) PeduncleColor Closest to 144B (Strong Yellow Green) with specks of very pale183A (Dark Red) throughout Petal Pubescence None Petal Length Lateral:Approximately 2.5 cm Upper: Approximately 3.0 cm Lower: Approximately2.5 cm Petal Width Lateral: Approximately 2.2 cm Upper: Approximately3.1 cm Lower: Approximately 3.8 cm Petal Shape Obcordate Petal ApexEmarginate (cleaved) Petal margin Entire Petal Base Attenuate PetalColor - Upper: Closest to but lighter than N75C (Pale Purple) with a mixof N78A (Strong Reddish Purple) and NN78A (Strong Reddish Purple) oneach petal, together forming a star-like shape. Petal Color - Lower:Closest to N75C with midveins of N79C (Dark Purplish Red) Petal Color -Eye Zone: Spot 60A (Deep Red) with background of 69C (Very Pale Purple)Spur Spur Color: Background of N155B (Pinkish White), 187A (Dark Red) atthe top tip, specks of 72B (Strong Reddish Purple) on the top half; 145C(Light yellow green) at end and an end tip of 144C (Strong Yellow Green)Spur Shape: Tubular, curved downward Spur Length: Approximately 7.4 cmSpur Diameter: Approximately 1.5 mm Stamens Stamen Form: Fused; splitinto 4 lobes Stamen Number: 1 Anther Length: 5.0 mm Anther Color: 64A(Moderate Purplish Red) Pistils Pistil Number: 1 Pistil Length 5.0 mmStigma Color: Mostly transparent 183A (Dark Red) Style Color: Backgroundcolor of 144A (Strong Yellow Green) with 183A (Dark Red) at top OvaryArrangement: Parietal Ovary Surface Color: 150A (Brilliant Yellow Green)Pollen & Seeds Pollen Amount: Abundant Pollen Color NN155A (YellowishWhite) Pollen Description Powdery Seed Production: None Observed

SAKIMP069 is most similar to the commercial Impatiens variety‘SAKIMP058’ (U.S. Pat. No. 10,638,692), however, there are differencesas listed in the table below.

TABLE 2 COMPARISON OF SAKIMP069 WITH ‘SAKIMP058’ SAKIMP069 SAKIMP058Flower Color, Closest to but lighter than Closest to RHS 75A Upper N75C(Pale Purple) with a mix (Light Purple) on the of N78A (Strong Reddishedges with RHS N74A Purple) and NN78A (Strong (Vivid Reddish Purple)Reddish Purple) on each petal, towards the center together forming astar-like shape. Plant Habit Compact; Mounding Vigorous; spreading

TABLE 3 COMPARISON OF SAKIMP069 WITH PARENTAL LINES Female Parent MaleParent SAKIMP069 NJ-1174A NJ-333-49H-6H Flower Pale Purple with LilacRose Color Reddish Purple Plant Compact; More compact More compact Habitmounding than SAKIMP069 than SAKIMP069

FURTHER EMBODIMENTS

Breeding with New Guinea Impatiens SAKIMP069

The goal of ornamental plant breeding is to develop new, unique andsuperior ornamental plants. The breeder initially selects and crossestwo or more parental lines, followed by repeated selfing and selection,producing many new genetic combinations. The breeder can theoreticallygenerate billions of different genetic combinations via crossing,selection, selfing and mutations. Therefore, a breeder will neverdevelop the same genetic variety, having the same traits from the exactsame parents.

Each year, the plant breeder selects the germplasm to advance to thenext generation. This germplasm is grown under unique and differentgeographical climatic and soil conditions and further selections arethen made during and at the end of the growing season. The varietiesthat are developed are unpredictable because the breeder's selectionoccurs in unique environments with no control at the DNA level, and withmillions of different possible genetic combinations being generated. Abreeder of ordinary skill in the art cannot predict the final resultinglines he develops, except possibly in a very gross and general fashion.The same breeder cannot produce the same variety twice by using the sameoriginal parents and the same selection techniques. Thisunpredictability results in the expenditure of large amounts of researchmonies to develop superior impatiens varieties.

Breeding programs combine desirable traits from two or more varieties orvarious broad-based sources into breeding pools from which varieties aredeveloped by selfing and selection of desired phenotypes. Pedigreebreeding is used commonly for the improvement of self-pollinatingplants. Two parents that possess favorable, complementary traits arecrossed to produce an F₁. An F₂ population is produced by selfing one orseveral F₁s. Selection of the best individuals may begin in the F₂population; then, beginning in the F₃, the best individuals in the bestfamilies are selected. Replicated testing of families can begin in theF₄ generation to improve the effectiveness of selection for traits withlow heritability. At an advanced stage of inbreeding (i.e., F₆ and F₇),the best lines or mixtures of phenotypically similar lines are testedfor potential release as new varieties.

Using New Guinea Impatiens SAKIMP069 to Develop Other Plants

SAKIMP069 plants can also provide a source of breeding material that maybe used to develop new impatiens plants and varieties. Plant breedingtechniques known in the art and used in an impatiens plant breedingprogram include, but are not limited to, recurrent selection, massselection, bulk selection, hybridization, mass selection, backcrossing,pedigree breeding, open-pollination breeding, restriction fragmentlength polymorphism enhanced selection, genetic marker enhancedselection, making double haploids, mutagenesis and transformation. Oftencombinations of these techniques are used. The development of impatiensvarieties in a plant breeding program requires, in general, thedevelopment and evaluation of homozygous varieties. There are manyanalytical methods available to evaluate a new variety. The oldest andmost traditional method of analysis is the observation of phenotypictraits, but genotypic analysis may also be used.

Additional Breeding Methods

Any plants produced using the SAKIMP069 plants disclosed in the presentapplication as at least one parent are also an embodiment. These methodsare well-known in the art and some of the more commonly used breedingmethods are described herein. Descriptions of breeding methods can befound in one of several reference books (e.g., Allard, “Principles ofPlant Breeding” (1999); and Vainstein, “Breeding for Ornamentals:Classical and Molecular Approaches,” Kluwer Academic Publishers (2002);Callaway, “Breeding Ornamental Plants,” Timber Press (2000).

Breeding steps that may be used in the New Guinea Impatiens SAKIMP069plant breeding program can include for example, pedigree breeding,backcrossing, mutation breeding, and recurrent selection. In conjunctionwith these steps, techniques such as RFLP-enhanced selection, geneticmarker enhanced selection (for example, SSR markers), Gene Editing andthe making of double haploids may be utilized.

As used herein, the term “plant” includes plant cells, plantprotoplasts, plant cell tissue cultures from which SAKIMP069 plants canbe regenerated, plant calli, plant clumps, and plant cells that areintact in plants or parts of plants, such as pollen, ovules, embryos,protoplasts, meristematic cells, callus, leaves, anthers, cotyledons,hypocotyl, pistils, roots, root tips, seeds, flowers, petiole, shoot, orstems and the like.

Pedigree Breeding

Pedigree breeding starts with the crossing of two genotypes, such as NewGuinea Impatiens SAKIMP069 and another different impatiens plant havingone or more desirable characteristics that is lacking or whichcomplements the New Guinea Impatiens SAKIMP069 phenotype. If the twooriginal parents do not provide all the desired characteristics, othersources can be included in the breeding population. In the pedigreemethod, superior plants are selfed and selected in successive filialgenerations. In the succeeding filial generations, the heterozygouscondition gives way to homogeneous varieties as a result ofself-pollination and selection. Typically, in the pedigree method ofbreeding, five or more successive filial generations of selfing andselection is practiced: F₁ to F₂; F₂ to F₃; F₃ to F₄; F₄ to F₅; etc.After a sufficient amount of inbreeding, successive filial generationswill serve to increase seed of the developed variety. Preferably, thedeveloped variety comprises homozygous alleles at about 95% or more ofits loci.

Backcross Breeding

Backcross breeding has been used to transfer genes for a simplyinherited, highly heritable trait into a desirable homozygous variety orinbred line which is the recurrent parent. The source of the trait to betransferred is called the donor parent. After the initial cross,individuals possessing the phenotype of the donor parent are selectedand repeatedly crossed (backcrossed) to the recurrent parent. Theresulting plant is expected to have the attributes of the recurrentparent and the desirable trait transferred from the donor parent. Thisis also known as single gene conversion and/or backcross conversion.

In addition to being used to create a backcross conversion, backcrossingcan also be used in combination with pedigree breeding. As discussedpreviously, backcrossing can be used to transfer one or morespecifically desirable traits from one variety, the donor parent, to adeveloped variety called the recurrent parent, which has overall goodcommercial characteristics yet lacks that desirable trait or traits.However, the same procedure can be used to move the progeny toward thegenotype of the recurrent parent, but at the same time retain manycomponents of the nonrecurrent parent by stopping the backcrossing at anearly stage and proceeding with selfing and selection. For example, aNew Guinea Impatiens plant may be crossed with another variety toproduce a first generation progeny plant. The first generation progenyplant may then be backcrossed to one of its parent varieties to create aBC₁ or BC₂. Progeny are selfed and selected so that the newly developedvariety has many of the attributes of the recurrent parent and yetseveral of the desired attributes of the nonrecurrent parent. Thisapproach leverages the value and strengths of the recurrent parent foruse in new New Guinea Impatiens varieties.

Therefore, another embodiment is a method of making a backcrossconversion of New Guinea Impatiens SAKIMP069, comprising the steps ofcrossing New Guinea Impatiens SAKIMP069 with a donor plant comprising adesired trait, selecting an F₁ progeny plant comprising the desiredtrait, and backcrossing the selected F₁ progeny plant to New GuineaImpatiens SAKIMP069. This method may further comprise the step ofobtaining a molecular marker profile of New Guinea Impatiens SAKIMP069and using the molecular marker profile to select for a progeny plantwith the desired trait and the molecular marker profile of New GuineaImpatiens SAKIMP069.

Recurrent Selection and Mass Selection

Recurrent selection is a method used in a plant breeding program toimprove a population of plants. New Guinea Impatiens SAKIMP069 issuitable for use in a recurrent selection program. The method entailsindividual plants cross-pollinating with each other to form progeny. Theprogeny are grown and the superior progeny selected by any number ofselection methods, which include individual plant, half-sib progeny,full-sib progeny, and selfed progeny. The selected progeny arecross-pollinated with each other to form progeny for another population.This population is planted and again superior plants are selected tocross-pollinate with each other. Recurrent selection is a cyclicalprocess and therefore can be repeated as many times as desired. Theobjective of recurrent selection is to improve the traits of apopulation. The improved population can then be used as a source ofbreeding material to obtain new varieties for commercial or breedinguse, including the production of a synthetic variety. A syntheticvariety is the resultant progeny formed by the intercrossing of severalselected varieties.

Mass selection is a useful technique when used in conjunction withmolecular marker enhanced selection. In mass selection, seeds fromindividuals are selected based on phenotype or selection requiresgrowing a population of plants in a bulk plot, allowing the plants toself-pollinate, harvesting the seed in bulk, and then using a sample ofthe seed harvested in bulk to plant the next generation. Also, insteadof self-pollination, directed pollination could be used as part of thebreeding program.

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating plants. A genetically variablepopulation of heterozygous individuals is either identified, or created,by intercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued.

Protoplast Fusion

Also known as somatic fusion, this process can be used with SAKIMP069 tocreate hybrids. The resulting hybrid plants have the chromosomes of eachparent and thus the process is useful for incorporating new traits. Theprotoplast fusion technique is well known in the art; see for exampleHamill J. D., Cocking E. C. (1988) Somatic Hybridization of Plants andits Use in Agriculture. In: Pais M. S. S., Mavituna F., Novais J. M.(eds) Plant Cell Biotechnology. NATO ASI Series (Series H: CellBiology), vol 18.

Mutation Breeding

Mutation breeding is another method of introducing new traits into NewGuinea Impatiens SAKIMP069. Mutations that occur spontaneously or areartificially induced can be useful sources of variability for a plantbreeder. The goal of artificial mutagenesis is to increase the rate ofmutation for a desired characteristic. Mutation rates can be increasedby many different means including temperature, long-term seed storage,tissue culture conditions, ionizing radiation, such as X-rays, Gammarays (e.g., cobalt 60 or cesium 137), neutrons, (product of nuclearfission by uranium 235 in an atomic reactor), Beta radiation (emittedfrom radioisotopes such as phosphorus 32 or carbon 14), or ultravioletradiation (preferably from 2500 to 2900 nm); chemical mutagens (such asbase analogues (5-bromo-uracil)), related compounds (8-ethoxy caffeine),antibiotics (streptonigrin), alkylating agents (sulfur mustards,nitrogen mustards, epoxides, ethylenamines, sulfates, sulfonates such asethyl methanesulfonate, sulfones, lactones), sodium azide,hydroxylamine, nitrous acid, methylnitrilsourea, or acridines; TILLING(targeting induced local lesions in genomes), where mutation is inducedby chemical mutagens and mutagenesis is accompanies by the isolation ofchromosomal DNA from every mutated plant line or seed and screening ofthe population of the seed or plants is performed at the DNA level usingadvanced molecular techniques. Once a desired trait is observed throughmutagenesis the trait may then be incorporated into existing germplasmby traditional breeding techniques. Details of mutation breeding can befound in Vainstein, “Breeding for Ornamentals: Classical and MolecularApproaches,” Kluwer Academic Publishers (2002); Sikora, Per, et al.,“Mutagenesis as a Tool in Plant Genetics, Functional Genomics, andBreeding” International Journal of Plant Genomics. 2011 (2011); 13pages. In addition, mutations created in other New Guinea Impatiensplants may be used to produce a backcross conversion of New GuineaImpatiens that comprises such mutation.

Gene Editing Using CRISPR

Targeted gene editing can be done using CRISPR/Cas9 technology (Saunders& Joung, Nature Biotechnology, 32, 347-355, 2014). CRISPR is a type ofgenome editing system that stands for Clustered Regularly InterspacedShort Palindromic Repeats. This system and CRISPR-associated (Cas) genesenable organisms, such as select bacteria and archaea, to respond to andeliminate invading genetic material. Ishino, Y., et al. J. Bacteriol.169, 5429-5433 (1987). These repeats were known as early as the 1980s inE. coli, but Barrangou and colleagues demonstrated that S. thermophiluscan acquire resistance against a bacteriophage by integrating a fragmentof a genome of an infectious virus into its CRISPR locus. Barrangou, R.,et al. Science 315, 1709-1712 (2007). Many plants have already beenmodified using the CRISPR system. See for example, U.S. ApplicationPublication No. WO2014068346 (György et al., Identification of aXanthomonas euvesicatoria resistance gene from pepper (Capsicum annuum)and method for generating plants with resistance); Martinelli, F. etal., “Proposal of a Genome Editing System for Genetic Resistance toTomato Spotted Wilt Virus” American Journal of Applied Sciences 2014;Noman, A. et al., “CRISPR-Cas9: Tool for Qualitative and QuantitativePlant Genome Editing” Frontiers in Plant Science Vol. 7 Nov. 2016; and“Exploiting the CRISPR/Cas9 System for Targeted Genome Mutagenesis inPetunia” Science Reports Volume 6: February 2016.

Gene editing can also be done using crRNA-guided surveillance systemsfor gene editing. Additional information about crRNA-guided surveillancecomplex systems for gene editing can be found in the followingdocuments, which are incorporated by reference in their entirety: U.S.Application Publication No. 2010/0076057 (Sontheimer et al., Target DNAInterference with crRNA); U.S. Application Publication No. 2014/0179006(Feng, CRISPR-CAS Component Systems, Methods, and Compositions forSequence Manipulation); U.S. Application Publication No. 2014/0294773(Brouns et al., Modified Cascade Ribonucleoproteins and Uses Thereof);Sorek et al., Annu. Rev. Biochem. 82:273-266, 2013; and Wang, S. et al.,Plant Cell Rep (2015) 34: 1473-1476. Therefore, it is another embodimentto use the CRISPR system on New Guinea Impatiens SAKI1V11³069 to modifytraits and resistances or tolerances to pests, herbicides, diseases, andviruses.

Gene Editing Using TALENs

Transcription activator-like effector nucleases (TALENs) have beensuccessfully used to introduce targeted mutations via repair of doublestranded breaks (DSBs) either through non-homologous end joining (NHEJ),or by homology-directed repair (HDR) and homology-independent repair inthe presence of a donor template. Thus, TALENs are another mechanism fortargeted genome editing using SAKIMP069. The technique is well known inthe art; see for example Malzahn, Aimee et al. “Plant genome editingwith TALEN and CRISPR” Cell & bioscience vol. 7 21. 24 Apr. 2017.

Therefore, it is another embodiment to use the TALENs system on NewGuinea Impatiens variety SAKIMP069 to modify traits and resistances ortolerances to pests, herbicides, and viruses.

Other Methods of Genome Editing

In addition to CRISPR and TALENs, two other types of engineerednucleases can be used for genome editing: engineered homingendonucleases/meganucleases (EMNs), and zinc finger nucleases (ZFNs).These methods are well known in the art. See for example, Petilino,Joseph F. “Genome editing in plants via designed zinc finger nucleases”In Vitro Cell Dev Biol Plant. 51(1): pp. 1-8 (2015); and Daboussi,Fayza, et al. “Engineering Meganuclease for Precise Plant GenomeModification” in Advances in New Technology for Targeted Modification ofPlant Genomes. Springer Science+Business. pp 21-38 (2015).

Therefore, it is another embodiment to use engineered nucleases on NewGuinea Impatiens variety SAKIMP069 to modify traits and resistances ortolerances to pests, herbicides, and viruses.

Additional Methods of Transformation

Additional methods include, but are not limited to, expression vectorsintroduced into plant tissues using a direct gene transfer method, suchas microprojectile-mediated delivery, DNA injection, electroporation,and the like. More preferably, expression vectors are introduced intoplant tissues by using either microprojectile-mediated delivery with abiolistic device or by using Agrobacterium-mediated transformation.Transformant plants obtained with the protoplasm of the subject NewGuinea Impatiens SAKIMP069 plants are intended to be within the scope ofthe embodiments of the application.

Single-Gene Conversions

When the term New Guinea Impatiens SAKIMP069 plant is used in thecontext of an embodiment of the present application, this also includesany single gene conversions of New Guinea Impatiens SAKIMP069. The termsingle gene converted plant as used herein refers to those impatiensplants which are developed by a plant breeding technique calledbackcrossing wherein essentially all of the desired morphological andphysiological characteristics of a variety are recovered in addition tothe single gene transferred into the variety via the backcrossingtechnique. Backcrossing methods can be used with one embodiment of thepresent application to improve or introduce a characteristic into thevariety. The term “backcrossing” as used herein refers to the repeatedcrossing of a hybrid progeny back to the recurrent parent, i.e.,backcrossing 1, 2, 3, 4, 5, 6, 7, 8, or more times to the recurrentparent. The parental New Guinea Impatiens plant that contributes thegene for the desired characteristic is termed the nonrecurrent or donorparent. This terminology refers to the fact that the nonrecurrent parentis used one time in the backcross protocol and therefore does not recur.The parental New Guinea Impatiens plant to which the gene or genes fromthe nonrecurrent parent are transferred is known as the recurrent parentas it is used for several rounds in the backcrossing protocol (Poehlman& Sleper (1994). In a typical backcross protocol, the original varietyof interest (recurrent parent) is crossed to a second variety(nonrecurrent parent) that carries the single gene of interest to betransferred. The resulting progeny from this cross are then crossedagain to the recurrent parent and the process is repeated until a NewGuinea Impatiens plant is obtained wherein essentially all of thedesired morphological and physiological characteristics of the recurrentparent are recovered in the converted plant, in addition to the singletransferred gene from the nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single gene of the recurrent variety ismodified or substituted with the desired gene from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphologicalconstitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross; one ofthe major purposes is to add some commercially important trait or traitsto the plant. The exact backcrossing protocol will depend on thecharacteristic or trait being altered to determine an appropriatetesting protocol. Although backcrossing methods are simplified when thecharacteristic being transferred is a dominant allele, a recessiveallele may also be transferred. In this instance, it may be necessary tointroduce a test of the progeny to determine if the desiredcharacteristic has been successfully transferred.

Many single gene traits have been identified that are not regularlyselected for in the development of a new variety but that can beimproved by backcrossing techniques. These traits are well-known in theart.

Introduction of a new trait or locus into New Guinea Impatiens SAKIMP069

New Guinea Impatiens SAKMP069 represents a new base of genetics intowhich a new locus or trait may be introgressed. Direct transformationand backcrossing represent two important methods that can be used toaccomplish such an introgression. The term backcross conversion andsingle locus conversion are used interchangeably to designate theproduct of a backcrossing program.

Backcross Conversions of New Guinea Impatiens SAKIMP069

A backcross conversion of New Guinea Impatiens SAKIMP069 occurs when DNAsequences are introduced through backcrossing (Allard, “Principles ofPlant Breeding” (1999) with New Guinea Impatiens SAKIMP069 utilized asthe recurrent parent. Both naturally occurring and transgenic DNAsequences may be introduced through backcrossing techniques. A backcrossconversion may produce a plant with a trait or locus conversion in atleast two or more backcrosses, including at least 2 crosses, at least 3crosses, at least 4 crosses, at least 5 crosses, and the like. Molecularmarker assisted breeding or selection may be utilized to reduce thenumber of backcrosses necessary to achieve the backcross conversion. Forexample, see, Openshaw, S. J., et al., Marker-assisted Selection inBackcross Breeding, Proceedings Symposium of the Analysis of MolecularData, Crop Science Society of America, Corvallis, Oreg. (August 1994),where it is demonstrated that a backcross conversion can be made in asfew as two backcrosses.

The complexity of the backcross conversion method depends on the type oftrait being transferred (single genes or closely linked genes ascompared to unlinked genes), the level of expression of the trait, thetype of inheritance (cytoplasmic or nuclear), and the types of parentsincluded in the cross. It is understood by those of ordinary skill inthe art that for single gene traits that are relatively easy toclassify, the backcross method is effective and relatively easy tomanage. See, Allard, “Principles of Plant Breeding” (1999). Desiredtraits that may be transferred through backcross conversion include, butare not limited to, sterility (nuclear and cytoplasmic), fertilityrestoration, drought tolerance, nitrogen utilization, ornamentalfeatures, disease resistance (bacterial, fungal, or viral), insectresistance, and herbicide resistance. In addition, an introgression siteitself, such as an FRT site, Lox site, or other site specificintegration site, may be inserted by backcrossing and utilized fordirect insertion of one or more genes of interest into a specific plantvariety. In some embodiments, the number of loci that may be backcrossedinto New Guinea Impatiens SAKIMP069 is at least 1, 2, 3, 4, or 5, and/orno more than 6, 5, 4, 3, or 2. A single locus may contain severaltransgenes, such as a transgene for disease resistance that, in the sameexpression vector, also contains a transgene for herbicide resistance.The gene for herbicide resistance may be used as a selectable markerand/or as a phenotypic trait. A single locus conversion of site specificintegration system allows for the integration of multiple genes at theconverted loci.

The backcross conversion may result from either the transfer of adominant allele or a recessive allele. Selection of progeny containingthe trait of interest is accomplished by direct selection for a traitassociated with a dominant allele. Transgenes or genes transferred viabackcrossing typically function as a dominant single gene trait and arerelatively easy to classify. Selection of progeny for a trait that istransferred via a recessive allele requires growing and selfing thefirst backcross generation to determine which plants carry the recessivealleles. Recessive traits may require additional progeny testing insuccessive backcross generations to determine the presence of the locusof interest. The last backcross generation is usually selfed to givepure breeding progeny for the gene(s) being transferred, although abackcross conversion with a stably introgressed trait may also bemaintained by further backcrossing to the recurrent parent withselection for the converted trait.

In addition, the above process and other similar processes describedherein may be used to produce first generation progeny impatiens seed byadding a step at the end of the process that comprises crossing NewGuinea Impatiens SAKIMP069 with the introgressed trait or locus with adifferent plant and harvesting the resultant first generation progenyseed.

Molecular Techniques Using New Guinea Impatiens SAKIMP069

The advent of new molecular biological techniques has allowed theisolation and characterization of genetic elements with specificfunctions. Traditional plant breeding has principally been the source ofnew germplasm, however, advances in molecular technologies have allowedbreeders to provide varieties with novel and much wanted commercialattributes. Molecular techniques such as transformation are popular inbreeding ornamental plants and well-known in the art. See Vainstein,“Breeding for Ornamentals: Classical and Molecular Approaches,” KluwerAcademic Publishers (2002).

Breeding with Molecular Markers

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select for the genome of the recurrent parent and against thegenome of the donor parent. Using this procedure can minimize the amountof genome from the donor parent that remains in the selected plants. Itcan also be used to reduce the number of crosses back to the recurrentparent needed in a backcrossing program. The use of molecular markers inthe selection process is often called genetic marker enhanced selection.Molecular markers may also be used to identify and exclude certainsources of germplasm as parental varieties or ancestors of a plant byproviding a means of tracking genetic profiles through crosses.Molecular markers, which includes markers identified through the use oftechniques such as Isozyme Electrophoresis, Restriction Fragment LengthPolymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs),Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats(SSRs), and Single Nucleotide Polymorphisms (SNPs), may be used in plantbreeding methods utilizing New Guinea Impatiens SAKIMP069. SeeVainstein, “Breeding for Ornamentals: Classical and MolecularApproaches,” Kluwer Academic Publishers (2002).

One use of molecular markers is Quantitative Trait Loci (QTL) mapping.QTL mapping is the use of markers, which are known to be closely linkedto alleles that have measurable effects on a quantitative trait.Selection in the breeding process is based upon the accumulation ofmarkers linked to the positive effecting alleles and/or the eliminationof the markers linked to the negative effecting alleles from the plant'sgenome. See for example, Fletcher, Richard S., et al., “QTL analysis ofroot morphology, flowering time, and yield reveals trade-offs inresponse to drought in Brassica napus” Journal of Experimental Biology.66 (1): 245-256 (2014). QTL markers can also be used during the breedingprocess for the selection of qualitative traits. For example, markersclosely linked to alleles or markers containing sequences within theactual alleles of interest can be used to select plants that contain thealleles of interest during a backcrossing breeding program. The markerscan also be used to select for the genome of the recurrent parent andagainst the genome of the donor parent. Using this procedure canminimize the amount of genome from the donor parent that remains in theselected plants. It can also be used to reduce the number of crossesback to the recurrent parent needed in a backcrossing program. The useof molecular markers in the selection process is often called geneticmarker enhanced selection. Molecular markers may also be used toidentify and exclude certain sources of germplasm as parental varietiesor ancestors of a plant by providing a means of tracking geneticprofiles through crosses.

Production of Double Haploids

The production of double haploids can also be used for the developmentof plants with a homozygous phenotype in the breeding program. Forexample, a New Guinea Impatiens plant for which New Guinea ImpatiensSAKIMP069 is a parent can be used to produce double haploid plants.Double haploids are produced by the doubling of a set of chromosomes(1N) from a heterozygous plant to produce a completely homozygousindividual. This can be advantageous because the process omits thegenerations of selfing needed to obtain a homozygous plant from aheterozygous source. For example, see, Ferrie, Alison M. R., et al.,“Review of Doubled Haploidy Methodologies in Ornamental Species”Propagation of Ornamental Plants. 11(2): pp. 63-77 (2011).

Thus, an embodiment is a process for making a substantially homozygousNew Guinea Impatiens SAKIMP069 progeny plant by producing or obtaining aseed from the cross of New Guinea Impatiens SAKIMP069 and anotherimpatiens plant and applying double haploid methods to the F₁ seed or F₁plant or to any successive filial generation.

In particular, a process of making seed retaining the molecular markerprofile of New Guinea Impatiens SAKIMP069 is contemplated, such processcomprising obtaining or producing F₁ seed for which New Guinea ImpatiensSAKIMP069 is a parent, inducing doubled haploids to create progenywithout the occurrence of meiotic segregation, obtaining the molecularmarker profile of New Guinea Impatiens SAKIMP069, and selecting progenythat retain the molecular marker profile of New Guinea ImpatiensSAKIMP069.

Expression Vectors for New Guinea Impatiens Transformation: Marker Genes

Plant transformation involves the construction of an expression vectorwhich will function in plant cells. Such a vector comprises DNAcomprising a gene under control of, or operatively linked to, aregulatory element (for example, a promoter). Expression vectors includeat least one genetic marker operably linked to a regulatory element (forexample, a promoter) that allows transformed cells containing the markerto be either recovered by negative selection, i.e., inhibiting growth ofcells that do not contain the selectable marker gene, or by positiveselection, i.e., screening for the product encoded by the geneticmarker. Many commonly used selectable marker genes for planttransformation are well-known in the transformation arts, and include,for example, genes that code for enzymes that metabolically detoxify aselective chemical agent which may be an antibiotic or an herbicide, orgenes that encode an altered target which is insensitive to theinhibitor. A few positive selection methods are also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptII) gene which, when under thecontrol of plant regulatory signals, confers resistance to kanamycin.Another commonly used selectable marker gene is the hygromycinphosphotransferase gene which confers resistance to the antibiotichygromycin.

Selectable marker genes for plant transformation not of bacterial origininclude, for example, mouse dihydrofolate reductase, plant5-enolpyruvylshikimate-3-phosphate synthase, and plant acetolactatesynthase (Eichholtz, et al., Somatic Cell Mol. Genet., 13:67 (1987);Shah, et al., Science, 233:478 (1986); Charest, et al., Plant Cell Rep.,8:643 (1990)).

Another class of marker genes for plant transformation requiresscreening of presumptively transformed plant cells, rather than directgenetic selection of transformed cells, for resistance to a toxicsubstance such as an antibiotic. These genes are particularly useful toquantify or visualize the spatial pattern of expression of a gene inspecific tissues and are frequently referred to as reporter genesbecause they can be fused to a gene or gene regulatory sequence for theinvestigation of gene expression. Commonly used marker genes forscreening presumptively transformed cells include β-glucuronidase (GUS),β-galactosidase, luciferase, and chloramphenicol acetyltransferase(Jefferson, R. A., Plant Mol. Biol. Rep., 5:387 (1987); Teeri, et al.,EMBO J., 8:343 (1989); Koncz, et al., Proc. Natl. Acad. Sci. USA, 84:131(1987); DeBlock, et al., EMBO J., 3:1681 (1984)).

Expression Vectors for New Guinea Impatiens Transformation: Promoters

Genes included in expression vectors must be driven by a nucleotidesequence comprising a regulatory element (for example, a promoter).Several types of promoters are well known in the transformation arts asare other regulatory elements that can be used alone or in combinationwith promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred.”Promoters that initiate transcription only in a certain tissue arereferred to as “tissue-specific.”A “cell-type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter which is under environmental control. Examples ofenvironmental conditions that may affect transcription by induciblepromoters include anaerobic conditions or the presence of light.Tissue-specific, tissue-preferred, cell-type specific, and induciblepromoters constitute the class of “non-constitutive” promoters. A“constitutive” promoter is a promoter that is active under mostenvironmental conditions. Many types of promoters are well known in theart.

Signal Sequences for Targeting Proteins to Subcellular Compartments

Transport of a protein produced by transgenes to a subcellularcompartment, such as the chloroplast, vacuole, peroxisome, glyoxysome,cell wall, or mitochondrion, or for secretion into the apoplast, isaccomplished by means of operably linking the nucleotide sequenceencoding a signal sequence to the 5′ and/or 3′ region of a gene encodingthe protein of interest. Targeting sequences at the 5′ and/or 3′ end ofthe structural gene may determine during protein synthesis andprocessing where the encoded protein is ultimately compartmentalized.Many signal sequences are well-known in the art. See, for example,Becker, et al., Plant Mol. Biol., 20:49 (1992); Knox, C., et al., PlantMol. Biol., 9:3-17 (1987); Lerner, et al., Plant Physiol., 91:124-129(1989); Frontes, et al., Plant Cell, 3:483-496 (1991); Matsuoka, et al.,Proc. Natl. Acad. Sci., 88:834 (1991); Gould, et al., J. Cell. Biol.,108:1657 (1989); Creissen, et al., Plant J., 2:129 (1991); Kalderon, etal., Cell, 39:499-509 (1984); Steifel, et al., Plant Cell, 2:785-793(1990).

Foreign Protein Genes: Transformation

Various promoters, targeting sequences, enhancing sequences, and otherDNA sequences can be inserted into the genome for the purpose ofaltering the expression of genes

Gene Silencing and Altering Gene Expression

Many techniques for altering gene expression are well-known to one ofskill in the art, including, but not limited to, knock-outs (such as byinsertion of a transposable element such as Mu (Vicki Chandler, TheMaize Handbook, Ch. 118 (Springer-Verlag 1994)) or other geneticelements such as a FRT, Lox, or other site specific integration sites;antisense technology (see, e.g., Sheehy, et al., PNAS USA, 85:8805-8809(1988) and U.S. Pat. Nos. 5,107,065, 5,453,566, and 5,759,829);co-suppression (e.g., Taylor, Plant Cell, 9:1245 (1997); Jorgensen,Trends Biotech., 8(12):340-344 (1990); Flavell, PNAS USA, 91:3490-3496(1994); Finnegan, et al., Bio/Technology, 12:883-888 (1994); Neuhuber,et al., Mol. Gen. Genet., 244:230-241 (1994)); RNA interference (Napoli,et al., Plant Cell, 2:279-289 (1990); U.S. Pat. No. 5,034,323; Sharp,Genes Dev., 13:139-141 (1999); Zamore, et al., Cell, 101:25-33 (2000);Montgomery, et al., PNAS USA, 95:15502-15507 (1998)), virus-induced genesilencing (Burton, et al., Plant Cell, 12:691-705 (2000); Baulcombe,Curr. Op. Plant Bio., 2:109-113 (1999)); target-RNA-specific ribozymes(Haseloff, et al., Nature, 334:585-591 (1988)); hairpin structures(Smith, et al., Nature, 407:319-320 (2000); U.S. Pat. Nos. 6,423,885,7,138,565, 6,753,139, and 7,713,715); MicroRNA (Aukerman & Sakai, PlantCell, 15:2730-2741 (2003)); ribozymes (Steinecke, et al., EMBO J.,11:1525 (1992); Perriman, et al., Antisense Res. Dev., 3:253 (1993));oligonucleotide mediated targeted modification (e.g., U.S. Pat. Nos.6,528,700 and 6,911,575); Zn-finger targeted molecules (e.g., U.S. Pat.Nos. 7,151,201, 6,453,242, 6,785,613, 7,177,766 and 7,788,044);transposable elements (e.g. Dubin, M. J., et al., Transposons: ablessing curse, Current opinion in plant biology, Vol: 42, Page: 23-29,2018 and Eric T. Johnson, Jesse B. Owens & Stefan Moisyadi (2016) Vastpotential for using the piggyBac transposon to engineer transgenicplants at specific genomic locations, Bioengineered, 7:1, 3-6); andother methods or combinations of the above methods known to those ofskill in the art.

Additional Transformation Embodiments

The foregoing methods for transformation may be used for producing atransgenic variety. The transgenic variety could then be crossed withanother (non-transformed or transformed) variety in order to produce anew transgenic variety. Alternatively, a genetic trait that has beenengineered into a particular New Guinea Impatiens line using theforegoing transformation techniques could be moved into another lineusing traditional backcrossing techniques that are well known in theplant breeding arts. For example, a backcrossing approach could be usedto move an engineered trait from a public, non-elite variety into anelite variety, or from a variety containing a foreign gene in its genomeinto a variety or varieties that do not contain that gene. As usedherein, “crossing” can refer to a simple x by y cross or the process ofbackcrossing depending on the context.

Likewise, by means of one embodiment, commercially important genes canbe expressed in transformed plants. More particularly, plants can begenetically engineered to express various phenotypes of commercialinterest, including, but not limited to, genes that confer resistance topests or disease, genes that confer resistance to an herbicide, genesthat confer or contribute to a value-added or desired trait, genes thatcontrol male sterility, genes that create a site for site specific DNAintegration, and genes that affect abiotic stress resistance. Manyhundreds if not thousands of different genes are known and couldpotentially be introduced into a New Guinea Impatiens plant according tothe invention. Non-limiting examples of particular genes andcorresponding phenotypes one may choose to introduce into a New GuineaImpatiens plant include one or more genes for insect tolerance, such asa Bacillus thuringiensis (Bt.) gene, pest tolerance such as genes forfungal disease control, herbicide tolerance such as genes conferringglyphosate tolerance, and genes for quality improvements such asenvironmental or stress tolerances, or any desirable changes in plantphysiology, growth, development, morphology or plant product(s). Forexample, structural genes would include any gene that confers insecttolerance including but not limited to a Bacillus insect control proteingene as described in WO 99/31248, herein incorporated by reference inits entirety, U.S. Pat. No. 5,689,052, herein incorporated by referencein its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, hereinincorporated by reference in their entirety. In another embodiment, thestructural gene can confer tolerance to the herbicide glyphosate asconferred by genes including, but not limited to Agrobacterium strainCP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat.No. 5,633,435, herein incorporated by reference in its entirety, orglyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No.5,463,175, herein incorporated by reference in its entirety.Alternatively, the DNA coding sequences can affect these phenotypes byencoding a non-translatable RNA molecule that causes the targetedinhibition of expression of an endogenous gene, for example viaantisense- or cosuppression-mediated mechanisms (see, for example, Birdet al., Biotech. Gen. Engin. Rev., 9:207, 1991). The RNA could also be acatalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desiredendogenous mRNA product (see for example, Gibson and Shillito, Mol.Biotech., 7:125, 1997). Thus, any gene which produces a protein or mRNAwhich expresses a phenotype or morphology change of interest is usefulfor the practice of one or more embodiments.

Tissue Culture

Further reproduction of the variety can occur by tissue culture andregeneration. Tissue culture of various tissues of ornamental plants andNew Guinea Impatiens SAKIMP069 and regeneration of plants therefrom iswell-known and widely published. For example, reference may be had to doValla Rego, Luciana et al., Crop Breeding and Applied Technology. 1(3):283-300 (2001); Komatsuda, T., et al., Crop Sci., 31:333-337 (1991);Stephens, P. A., et al., Theor. Appl. Genet., 82:633-635 (1991);Komatsuda, T., et al., Plant Cell, Tissue and Organ Culture, 28:103-113(1992); Dhir, S., et al., Plant Cell Reports, 11:285-289 (1992); Pandey,P., et al., Japan J. Breed., 42:1-5 (1992); and Shetty, K., et al.,Plant Science, 81:245-251 (1992). Thus, another embodiment is to providecells which upon growth and differentiation produce New Guinea Impatiensplants having the physiological and morphological characteristics of NewGuinea Impatiens SAKIMP069 described in the present application.

Regeneration refers to the development of a plant from tissue culture.The term “tissue culture” indicates a composition comprising isolatedcells of the same or a different type or a collection of such cellsorganized into parts of a plant. Exemplary types of tissue cultures areprotoplasts, calli, plant clumps, and plant cells that can generatetissue culture that are intact in plants or parts of plants, such aspollen, ovules, embryos, protoplasts, meristematic cells, callus,pollen, leaves, ovules, anthers, cotyledons, hypocotyl, pistils, roots,root tips, flowers, seeds, petiole, shoot, or stems, and the like. Meansfor preparing and maintaining plant tissue culture are well-known in theart.

While a number of exemplary aspects and embodiments have been discussedabove, those of skill in the art will recognize certain modifications,permutations, additions and sub-combinations thereof. It is thereforeintended that the following appended claims and claims hereafterintroduced are interpreted to include all such modifications,permutations, additions, and sub-combinations as are within their truespirit and scope.

One or more aspects may be embodied in other specific forms withoutdeparting from its spirit or essential characteristics. The describedembodiments are to be considered in all respects only as illustrativeand not restrictive. The scope of the embodiments is, therefore,indicated by the appended claims rather than by the foregoingdescription. All changes which come within the meaning and range ofequivalency of the claims are to be embraced within their scope. Theforegoing discussion of the embodiments has been presented for purposesof illustration and description. The foregoing is not intended to limitthe embodiments to the form or forms disclosed herein. In the foregoingDetailed Description for example, various features of the embodimentsare grouped together in one or more embodiments for the purpose ofstreamlining the disclosure. This method of disclosure is not to beinterpreted as reflecting an intention that the claimed embodimentsrequire more features than are expressly recited in each claim. Rather,as the following claims reflect, inventive aspects lie in less than allfeatures of a single foregoing disclosed embodiment. Thus, the followingclaims are hereby incorporated into this Detailed Description, with eachclaim standing on its own as a separate preferred embodiment.

Moreover, though the description of the embodiments has includeddescription of one or more embodiments and certain variations andmodifications, other variations and modifications are within the scopeof the embodiments (e.g., as may be within the skill and knowledge ofthose in the art, after understanding the present disclosure). It isintended to obtain rights which include alternative embodiments to theextent permitted, including alternate, interchangeable and/or equivalentstructures, functions, ranges or acts to those claimed, whether or notsuch alternate, interchangeable and/or equivalent structures, functions,ranges or acts are disclosed herein, and without intending to publiclydedicate any patentable subject matter.

The use of the terms “a,” “an,” and “the,” and similar referents in thecontext of describing the embodiments (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext. The terms “comprising,” “having,” “including,” and “containing”are to be construed as open-ended terms (i.e., meaning “including, butnot limited to,”) unless otherwise noted. Recitation of ranges of valuesherein are merely intended to serve as a shorthand method of referringindividually to each separate value falling within the range, unlessotherwise indicated herein, and each separate value is incorporated intothe specification as if it were individually recited herein. Forexample, if the range 10-15 is disclosed, then 11, 12, 13, and 14 arealso disclosed. All methods described herein can be performed in anysuitable order unless otherwise indicated herein or otherwise clearlycontradicted by context. The use of any and all examples, or exemplarylanguage (e.g., “such as”) provided herein, is intended merely to betterilluminate the embodiments and does not pose a limitation on the scopeof the embodiments unless otherwise claimed. No language in thespecification should be construed as indicating any non-claimed elementas essential to the practice one or more embodiments.

DEPOSIT INFORMATION

A deposit of the Sakata Seed Corporation proprietary New GuineaImpatiens variety SAKIMP069 plant tissue disclosed above and recited inthe appended claims has been made with the Provasoli-Guillard NationalCenter for Marine Algae and Microbiota, Bigelow Laboratory for OceanSciences (NCMA), 60 Bigelow Drive, East Boothbay, Me. 04544. The date ofdeposit was Apr. 22, 2022. The NCMA No. is 202204066. The deposit ofplant tissue was taken from the same deposit maintained by Sakata SeedCorporation since prior to the filing date of this application. Thedeposit will be maintained in the NCMA depository for a period of 30years, or 5 years after the most recent request, or for the enforceablelife of the patent, whichever is longer, and will be replaced ifnecessary, during that period. Upon issuance, all restrictions on theavailability to the public of the deposit will be irrevocably removedconsistent with all of the requirements of 37 C.F.R. §§ 1.801-1.809.

What is claimed is:
 1. A plant of New Guinea Impatiens varietySAKIMP069, wherein a representative sample of plant tissue of SAKIMP069was deposited under NCMA No.
 202204066. 2. A plant, or a plant partthereof produced by growing the plant of claim 1, wherein the plant orplant part comprises at least one cell of New Guinea Impatiens varietySAKIMP069.
 3. A New Guinea Impatiens plant, or part thereof, having allof the physiological and morphological characteristics of the New GuineaImpatiens plant of claim
 1. 4. A tissue or cell culture of regenerablecells produced from the plant of claim
 1. 5. The tissue or cell cultureof claim 4, comprising tissues or cells from a plant part selected fromthe group consisting of leaves, pollen, embryos, cotyledons, hypocotyl,meristematic cells, roots, root tips, pistils, anthers, flowers, andstems.
 6. A New Guinea Impatiens plant regenerated from the tissue orcell culture of claim 5, wherein said plant has all of the morphologicaland physiological characteristics of New Guinea Impatiens SAKIMP069. 7.A method of vegetatively propagating the plant of claim 1, comprisingthe steps of: collecting tissue or cells capable of being propagatedfrom said plant; cultivating said tissue or cells to obtain proliferatedshoots; and rooting said proliferated shoots to obtain rooted plantlets;or cultivating said tissue or cells to obtain proliferated shoots, or toobtain plantlets.
 8. A New Guinea Impatiens plant produced by growingthe plantlets or proliferated shoots produced by the method of claim 7.9. A method for producing an embryo or seed, wherein the methodcomprises crossing the plant of claim 1 with another plant andharvesting the resultant embryo or seed.
 10. A method of determining thegenotype of the New Guinea Impatiens plant of claim 1, wherein saidmethod comprises obtaining a sample of nucleic acids from said plant anddetecting in said nucleic acids a plurality of polymorphisms.
 11. Amethod of producing a New Guinea Impatiens plant resistant to the groupconsisting of herbicides, insects, and disease, wherein the methodcomprises transforming the New Guinea Impatiens plant of claim 1 with atransgene, and wherein said transgene confers resistance to anherbicide, insects, or disease.
 12. An herbicide, insect, or diseaseresistant plant produced by the method of claim
 11. 13. A method fordeveloping a New Guinea Impatiens plant in a plant breeding program,comprising applying plant breeding techniques comprising crossing,recurrent selection, mutation breeding, wherein said mutation breedingselects for a mutation that is spontaneously or naturally induced orartificially induced, backcrossing, pedigree breeding, marker enhancedselection, haploid/double haploid production, or transformation to aplant of New Guinea Impatiens variety SAKIMP069, wherein arepresentative sample of plant tissue of SAKIMP069 was deposited underNCMA No. 202204066, or its parts, wherein application of said techniquesresults in development of a New Guinea Impatiens plant.
 14. A method ofintroducing a mutation into the genome of New Guinea Impatiens plantSAKIMP069, said method comprising mutagenesis of the plant, or plantpart thereof, of claim 1, wherein the method is performed using amaterial or technique selected from the group consisting of temperature,long-term seed storage, tissue culture conditions, ionizing radiation,chemical mutagens, or targeting induced local lesions in genomes, andwherein the resulting plant comprises at least one genome mutation. 15.A method of editing the genome of New Guinea Impatiens plant SAKIMP069,wherein a representative sample of plant tissue of SAKIMP069 wasdeposited under NCMA No. 202204066, wherein said method wherein themethod is performed using a material or technique selected from thegroup consisting of zinc finger nucleases, transcription activator-likeeffector nucleases (TALENs), engineered homingendonucleases/meganucleases, and the clustered regularly interspacedshort palindromic repeat (CRISPR)-associated protein9 (Cas9) system. 16.A New Guinea Impatiens plant produced by the method of claim 15, whereinsaid plant has all of the morphological and physiologicalcharacteristics of New Guinea Impatiens SAKIMP069.
 17. A New GuineaImpatiens seed produced by growing the plant of claim
 1. 18. A method ofproducing a New Guinea Impatiens plant, or part thereof, produced bygrowing the seed of claim
 17. 19. A seed produced by the method of claim9, wherein the female parent is SAKIMP069.
 20. The method of claim 9,further comprising producing a plant, or a part thereof, from theresultant embryo or seed.